104. BIOINSTRUMENTATION
Unit –I
1. Principles of Microscopy – components of microscope – mechanical and optical; Resolving
power of different microscopes. Image magnification, contrast & resolution. Resolution
power and calculation, Numerical aperture, Refractive index, image illumination,
2. Type of microscopy‐Basic structure and function of‐ Light microscopy‐ Bright field, Dark
field, phase contrast, florescent, Confocal, Scanning & NDIC; Electron microscopes –
TEM, SEM, & Scanning Tunneling M.
3. Different methods of fixation and staining techniques of electron microscope. Microtomy:
Types, principal and applications. Freeze –etch and freeze fracture techniques.
4. Radio labeling techniques detection and measurement of different types of radio isotopes
used in biology and their application in biological science. Half life, Safety guidelines,
Molecular imaging of radioactive material.
Unit – II
1. Radio activity: Radio isotropic tracers – Methodology, Radio labeling techniques, Radio
metric analysis stable and radioactive isotopes, labeling, detection and measurement of
isotopes: Geign‐Muller counter, liquid scintillating counter and gamma counter.
2. Centrifugation‐ Principle, types and applications; centrifugation techniques‐ preparative
and analytical methods, density gradient centrifugation, Ultra centrifugation;
sedimentation coefficient and factor affecting centrifugation.
3. Principle, types and application of Electrophoresis – frontal & Zonal Electrophoresis, Paper,
agarose gel, PAGE, SDS‐PAGE, Capillary electrophoresis (CE).
4. Principles of ‐ Isoelectric focusing, 2D gel electrophoreses and Isotachophoresis, pulse field
Gel electrophoresis (PFGE); Application of electrophoresis in the diagnostic laboratory,
mutation detection.
Unit III
1. Principles, types and application of chromatography – Partition, adsorption, Gel
filiations Paper column, thin layer, Gas, Ion exchange and Molecular sieve; various
stationary and mobile phases, sample preparation, matrix materials, Column packing,
analyte development & elution.
2. Principle and application of affinity chromabrography‐ HPLC, FPLC (fast protein liquid
chromatography), Gas chromatography, Capillary electro chromatography, Hydroxy
apatite chromatography, Hydrophobic interaction chromatography.
3. Photometry: Law of absorption and radiation, principle, instrumentation and application of
UV‐visible spectrophotometry, infrared (IR) spectroscopy and Raman spectroscopy.
4. Atomic absorption spectroscopy: Principle instrumentation and application; Fluorescence
spectroscopy and Quenching.
Unit ‐IV
1. Electron spin resonance (ESR) spectroscopy and Nuclear Magnetic resonance (NMR):
Principle, Instrumentation and application.
2. Circular Dichrorism spectroscopy (CD): Principle, instrumentation and application.
3. Principle and applications of colorimetry, fluorescence flame photometry, fluorimetry,
polarimetry, turbidometry and Viscometry.
4. X‐ray crystallography.
Unit ‐ V
1. Mass spectrometry: components of mass spectrometer – vacuum system, ion source,
sample inlet, analyzer and detectors; Matrix.
2. Quadruople mass spectrometry, plasma desorption ionization, MALDI‐TOF, Peptide
mass determination and other application.
3. Surface plasma resonance methods and its applications.
4. Flow cytometry‐ FACS, MACS: Principle, instrumentation and application.
Practicals
1. Verification of Beer’s law
2. Determination of absorbtion maxima
3. Electrophoresis of Proteins‐ native and under denaturing conditions.
4. Aminoacid and carbohydrate separations by paper & thin layer chromatography
5. Gas chromatography
6. Ion exchange and gel filtration chromatography
7. Separation of subcellular organelles by differential centrifugation
8. Separation of blood cells by density gradient centrifugation
Books
1. Physical Biochemistry : Applications to Biochemistry and Molecular Biology by Freifelder
2. Biochemical Techniques : Theory and Practice by Robyt and White
3. Principles of Instrumental Analysis by Skoog and West
4. Analytical Biochemistry by Holme and Peck
5. Biological Spectroscopy by Campbell and Dwek
6. Organic Spectroscopy by Kemp
7. A Biologist’s Guide to Pronciples and Techniques of Practical Biochemistry
by Wilson and Goulding
8. Principles of Instrumental Analysis by Skoog, Hollar and Nicman
